Figure 7.
LRRK2 expression causes UPS disruption in zebrafish. (a) LRRK2 expression in zebrafish. Zebrafish embryos were injected with empty vector or WTLRRK2 at the 1–2 cell stage and lysed at 12 h.p.f. LRRK2 protein was detected by western blotting. Asterisk indicates unspecific band; arrow indicates LRRK2 band. (b) Zebrafish embryos at 12 h.p.f. following injection of UbG76V–GFP with either WTLRRK2 (I & IV) or empty vector (II & V). Uninjected siblings (III & VI) are included for comparison. Injection of WTLRRK2 (IV) or GSLRRK2 (not shown) resulted in a high and stable level of UbG76V–GFP expression. Typically, expression was observed in more than 30 cells. Injection of LRRK1 (not shown) or vector (V) resulted in a low level of UbG76V–GFP expression. In vector- and LRRK1-injected embryos, expression was observed in less than 10 cells per embryo. (c) Quantification of (b). Graph represents data of three independent experiments as mean percentage of fish with UbG76V–GFP-positive cells±S.D. Statistical analysis was performed by two-tailed, unpaired Student's t-test. **P<0.01, ***P<0.001, N=3. (d) Increased levels of UbG76V–GFP were detected in WTLRRK2-expressing zebrafish by western blot. Asterisk (*) indicates unspecific band; arrow indicates LRRK2 band